Mining for Active Molecules in Probiotic Supernatant by Combining Non-Targeted Metabolomics and Immunoregulation...

Area of Science:

• Microbiology and Immunology
• Metabolomics
• Biochemistry

Background:

• Chronic respiratory diseases like asthma are increasing globally, necessitating novel therapeutic strategies.
• Probiotics show potential in disease prevention and symptom reduction through immune system modulation via secreted metabolites.
• Previous research identified the amino acid d-Trp as having immunoregulatory bioactivity.

Purpose of the Study:

• To associate metabolic pathway activity with the in vitro immunomodulatory activity of probiotic strains.
• To identify key metabolic pathways involved in the bioactivity of probiotic supernatants.
• To explore the potential of metabolomics in discovering novel bioactive compounds from probiotics.

Main Methods:

• Metabolic footprinting of 25 commercial probiotic strains using untargeted liquid chromatography-mass spectrometry (LC-MS).
• Sample preparation involved protein precipitation and solid-phase extraction (SPE) with HLB and CN-E cartridges.
• Multivariate statistical analyses and pathway analysis were employed to distinguish bioactive from non-bioactive samples.

Main Results:

• Metabolomic analysis revealed an overrepresentation of the tryptophan metabolic pathway in bioactive probiotic supernatants.
• Specific molecular features within the tryptophan pathway were associated with observed in vitro immunomodulatory activity.
• The study successfully differentiated bioactive and non-bioactive probiotic samples based on metabolic profiles.

Conclusions:

• The tryptophan metabolic pathway is implicated in the immunomodulatory effects of certain probiotics.
• Metabolomics is a powerful tool for identifying bioactive compounds and driving product development in the probiotic industry.
• This research provides a foundation for discovering novel immunoregulatory molecules from probiotic sources.